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Device and method for isolating cells, bioparticles and/or molecules used for animal biotechnology (including biological research) and medical diagnosis from liquids

A technology of biotechnology and biological particles, applied in the direction of scientific instruments, instruments, biological tests, etc., can solve problems such as unsustainable operation

Inactive Publication Date: 2012-11-07
PLURISELECT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The disadvantage of all these sorting methods is that they do not work continuously, that is, blood or lymphocyte samples are collected and incubated with immuno-magnetic particles

Method used

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  • Device and method for isolating cells, bioparticles and/or molecules used for animal biotechnology (including biological research) and medical diagnosis from liquids
  • Device and method for isolating cells, bioparticles and/or molecules used for animal biotechnology (including biological research) and medical diagnosis from liquids
  • Device and method for isolating cells, bioparticles and/or molecules used for animal biotechnology (including biological research) and medical diagnosis from liquids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0026] Isolation of CD4 Positive Cells from Rat Whole Blood

[0027] Ascites antibody (RIB5 / 2) was done according to the protocol of Millipore Montage's Antibody Purification Kit (LSK2ABG20).

[0028] After purification, use SDS denaturing gel (10%) to detect

[0029] Conjugation of Anti-CD4 Antibody to Polypropylene (PMA) Particles

[0030] 1. Centrifuge 1ml of PMA (particle diameter=40μm+ / -10μm; 10mg / ml; COOH / PEG-COOH modification) for 2 minutes at 3,000xg, remove the supernatant and inhale in 1ml of 0.1M MES pH6.3 solution.

[0031] 2. Dissolve 2mg of EDC and 2.4mg of N-hydroxysuccinimide in 0.5ml of 0.1M MES pH6.3 buffer solution and add to the suspension of PMA particles. Incubate for 1 hour at room temperature with rotation (activation of particles).

[0032] 3. Separation of PMA particles: Centrifuge and elute twice with 0.1M MES r pH6.3 solution.

[0033] 4. Aspirate activated PMA particles into 1 ml of 0.1M MES pH6.3 buffer solution with 100-150 μg of antibody; co...

Embodiment approach 2

[0052] Protein Isolation (IgG) from Cell Lysates

[0053] Ascites antibody (RIB5 / 2) was done according to the protocol of Millipore Montage's Antibody Purification Kit (LSK2ABG20).

[0054] Purification was then checked with SDS denaturing gel (10%).

[0055] (Mouse IgG 2) Conjugation of Anti-CD4 Antibody to Polypropylene (PMA) Particles

[0056] 1. Centrifuge 1ml of PMA (particle diameter=40μm+ / -10μm; 10mg / ml; COOH / PEG-COOH modification) for 2 minutes at 3,000xg, remove the supernatant and inhale in 1ml of 0.1M MES pH6.3 buffer solution.

[0057] 2. 2mg of EDC and 2.4mg of N-hydroxysuccinimide buffered in 0.5ml of 0.1M MES pH6.3 buffer solution were added to the suspension of PMA particles. Incubate for 1 hour at room temperature with rotation (activation of particles).

[0058] 3. Separation of PMA particles: Centrifuge and elute twice with 0.1M MES r pH6.3 buffer solution.

[0059] 4. Aspirate active PMA particles into 1ml 0.1M MES pH6.3 buffer solution with 100-150μg ...

Embodiment approach 3

[0074] Figure 7-11 shows an example of the invention or a part thereof implemented on the basis of an instrument, Figure 7 Represents a typical system that can be used continuously. The dotted arrow indicates the hose system. The arrow tip marks the flow direction of the fluid. Starting from the organism (mammal), the body fluid, such as blood, is pumped or directed to a container containing functional particles. The reaction vessel, possibly through a blood pump or a vacuum tube (marked as a circle in Figure 8), interacts in admixture with the constituents of the body fluid. directing a medium of cell mixture and functional particles to a particle separation system, according to one example, the medium of cell mixture and functional particles is directed to a sieve and passed through a hollow fiber membrane, and then separated into a cell mixture free of functional particles and a medium containing Cell mixture of functional particles ( Figure 9 ). But the mixture witho...

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Abstract

The invention relates to a device and a method for identifying and isolating specific bioparticles and also dissolved biomolecules from liquids, with the aid of suitable carriers and known immobilisation techniques. The device can be used both intermittently and also for the direct and continuous treatment of liquids. The invention can be used with animals, in biotechnology, (including biologicalresearch) and medical diagnostics.

Description

technical field [0001] The present invention relates to a method and apparatus for separating cells, biological particles and / or molecules from a fluid. By means of the present invention and using suitable supports and known immobilization methods, specific biological particles can be identified and isolated. The invention has application in the fields of animals, biotechnology (including biological research) and medical diagnosis. Background technique [0002] Separation of cells, biological particles and / or molecules from fluids is important in many fields and many methods are known. [0003] Fluorescence-activated cell sorting (FACS) has been used for cell analysis and cell isolation for many years. This method is a superior method for the analysis of specific cell populations by surface markers. However, problems arise when using FACS to isolate large numbers of cells. Cell-containing media must be highly dilute, take a relatively long time to isolate a large number ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/48A61M1/36G01N37/00A61M1/38A61M1/34
CPCG01N2001/4016G01N33/5002A61M1/362A61M1/34G01N37/00
Inventor 汉斯-维尔纳·海因里希
Owner PLURISELECT
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